A dynamic interplay of enhancer elements regulates Klf4 expression in naïve pluripotency
Open Access
- 1 September 2017
- journal article
- Published by Cold Spring Harbor Laboratory in Genes & Development
- Vol. 31 (17), 1795-1808
- https://doi.org/10.1101/gad.303321.117
Abstract
Transcription factor (TF)-directed enhanceosome assembly constitutes a fundamental regulatory mechanism driving spatiotemporal gene expression programs during animal development. Despite decades of study, we know little about the dynamics or order of events animating TF assembly at cis-regulatory elements in living cells and the long-range molecular “dialog” between enhancers and promoters. Here, combining genetic, genomic, and imaging approaches, we characterize a complex long-range enhancer cluster governing Krüppel-like factor 4 (Klf4) expression in naïve pluripotency. Genome editing by CRISPR/Cas9 revealed that OCT4 and SOX2 safeguard an accessible chromatin neighborhood to assist the binding of other TFs/cofactors to the enhancer. Single-molecule live-cell imaging uncovered that two naïve pluripotency TFs, STAT3 and ESRRB, interrogate chromatin in a highly dynamic manner, in which SOX2 promotes ESRRB target search and chromatin-binding dynamics through a direct protein-tethering mechanism. Together, our results support a highly dynamic yet intrinsically ordered enhanceosome assembly to maintain the finely balanced transcription program underlying naïve pluripotency.Keywords
Funding Information
- California Institute of Regenerative Medicine (LA1-08013)
- National Institute of Arthritis and Musculoskeletal and Skin Diseases
- National Institutes of Health
- Howard Hughes Medical Institute
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